Method and composition for reducing plasma lipid levels

ABSTRACT

Plasma lipid levels in a mammal are reduced by compounds such as 4,4&#39;&#39;-methylenebis(2,6-di-tert-butylphenol). A preferred embodiment encompasses methods and compositions for reducing plasma lipid levels based on compounds having the formula   WHEREIN Ra and Rb are tertiary alkyl radicals, most preferably tertbutyl, R and Rc are alkyl radicals of one to four carbons, and Rd and Re are independently selected from hydrogen or methyl.   D R A W I N G

United States Patent [1 1 Lerner llll 3,873,726

[ Mar. 25, I975 METHOD AND COMPOSITION FOR REDUCING PLAsMA LIPID LEvELs [52] U.S. Cl. 424/347, 424/346 [51] Int. Cl A6lk 27/00 [5 8] Field of Search 424/346, 347

[56] References Cited UNITED STATES PATENTS 2,651,572 9/1953 Bickoff 99/8 3.529066 9/l970 Barnhart 424/346 FOREIGN PATENTS OR APPLICATIONS 1,001.351 4/l963 United Kingdom Primary Examiner-Sam Rosen Attorney, Agent, or Firm-D. L. Johnson; Robert A. Linn; Joseph D. Odenweller [57] ABSTRACT Plasma lipid levels in a mammal are reduced by compounds such as 4,4 -methylenebis(2,6-di-tertbutylphenol).

A preferred embodiment encompasses methods and compositions for reducing plasma lipid levels based on compounds having the formula a b R R Ed I HO e H R C R R wherein R and R are tertiary alkyl radicals, most preferably tert-butyl, R and R" are alkyl radicals of one to four carbons,

and R" and R" are independently selected from hydrogen or methyl.

23 Claims, N0 Drawings METHOD AND COMPOSITION FOR REDUCING PLASMA LIPID LEVELS BACKGROUND OF INVENTION This application is a continuation-in-part of application Ser. No. 705,295, filed Feb. 14, 1968, which in turn is a continuation-in-part of application Ser. No. 3l7,796, filed Oct. 21, 1963, both now abandoned.

In said earlier application Ser. No. 317,796, I disclosed methods and compositions (for reducing serum lipid levels) based on compounds having the formula (mm i e wherein R and R are each selected from the group consisting of hydrogen and hydrocarbon radicals,

R is a hydrocarbon radical and n and p are integers of from -4, such that when n and p are each at least I, the radicals collectively represented by R may, on each phenyl moiety, be the same or different.

The hydrocarbon radicals are preferably alkyl of from l-l2 carbon atoms, cycloalkyl of from 6 to l2 carbon atoms, aryl of from 6-12 carbon atoms and aralkyl of from 7-12 carbon atoms.

SUMMARY OF THE INVENTION This application, like said earlier application Ser. No. 705,295, is directed to an embodiment of the invention described within the aforementioned parent application; to wit, methods and compositions for reducing plasma lipid levels which are based on lipid lowering compounds having the formula 2a R H 4 R 1 g (I) Y wherein R and R are lower alkyl radicals (that is, alkyl radicals of l to 8 carbon atoms), Y and Y are hydrogen or lower alkyl radicals, and R and R is no more than about 5.

DESCRIPTION OF PREFERRED EMBODIMENTS A preferred embodiment encompasses methods and compositions for reducing plasma lipid levels based on compounds having the formula a b R R I HO- Ci! OH R R R III wherein R and R" are tertiary alkyl radicals, most preferably tert-butyl,

R and R are alkyl radicals of one to four carbons,

and

R and R" are independently selected from hydrogen or methyl.

A preferred compound is 4,4-methylenebis(2,6-ditert-butylphenol). That compound is also known as di- C3,5-di-tert-butyl-4-hydroxyphenyl)methane. Other compounds are considered full equivalents. These are illustrated by the following non-limiting list:

A skilled practitioner can readily see that compounds closely related to those named above are within the scope of this invention. Such compounds include phenols substituted with other alkyl groups, haloalkyl, such as trifluoromethyl, cycloalkyl, halogen, and the like. In addition, closely related compounds within the scope of this invention include those with benzyl radical substitution. Such compounds are illustrated by:

4,4'-methylenebis-[ 2,6-bis( a-methylbenzyl )phenol] 4,4'-methylenebis-[ 2-methyl-6-( amethylbenzyl )phenol] 4,4'-isopropylidenebis-[ 2-tert-butyl-6-( amethylbenzyl )-phenol] and the like.

In addition, positional isomers of the above compounds are within the scope of this invention. Such compounds are illustrated by 2,2-methylenebis-(6- tert-butyl-p-cresol) and 2,2-methylenebis-(6-tertbutyli chlorophenol).

Furthermore, compounds not so heavily substituted are within the scope ofthis invention, Such compounds are illustrated by 2-tert-butyI-4,4-methylenediphenol. In addition, the solubility of the active phenols can be altered by modifying the phenolic moiety. This is illustrated by esters and ethers of the phenols such as bis( 3- ,5-di-tert-butyl-4-ethoxyphenyl)-methane.

The Compounds of this invention can be prepared by methods well known in the art. Those di-(hydroxyphenyl)methanes in which the phenyl moieties are the same can be prepared by condensation of an approprimagnesium sulfate, and the volatiles removed on a rotary evaporator. The heavy oil which remained was found to crystallize when treated with isopropanol. Two recrystallizations of these crystals from isopropaate phenol with an appropriate aldehyde, in the pres- 5 nol gave 5.34 g 22.3 per cent) of white crystals which ence of base, as described in U.S. Pat. No. 2,807,653. melted at ll1 1l3C The VPC of this material Where the methylene bridge is disubstituted, a ketone h w d l a i or i i y, Th i frared e trum can be used. U.S. Pat. No. 2,829,175 describes a did t h an OH stretching between 33-3800 method whereby anhydrous i'ici Zinc Chloride can and was consistent with the proposed structure of the be used to condense 2-, 2,4-, and 3-hydrocarbyl SubSfil0 diethyl ether of the starting phenol. Anal: Calcd. for tuted phenols with an appropriate ketone. These meth- C H O C, 82,44; H, 10,90, m l wt, 480.75, ods can also be used to prepare those di-(hydroxy- F d; C 825; H 110, m; r 489 phehyiimethahes in which the P y moieties are The compounds of this invention are remarkably efferent. In these cases two different phenols are confective in brin ing about a rapid drop in circulating densed with an appropriate aldehyde or ketone, using serum lipid levels using very low dosage regimens. This an appropriate condensation procedure, and ordinary allows use in pharmaceutical forms both convenient for separation procedures, such as fractional distillation, administration as well as pleasant for patient consumpcrystallization and solvent extraction procedures, etc. tion. can be used. To demonstrate the outstanding effectiveness of the The compound, bi 3,5-di-t t-b t l-4- compounds of this invention tests were performed on ethoxyphenyl)-methane which was mentioned above 26 g g which are Separated ihlo four was made by the following procedure; groups. The Control GI'OUP Of 3 male and 3 female D di h d id was prepared f commercial dogs was maintained on a diet of Allied Mills Tail sodium hydride in mineral oil. The mineral oil was re- Wagger Dog Food-Kruihs to which was added moved by filtration under nitrogen and the solid was Weight P Gem of Wesson in Experimental Groups h d several times i h d petroleum ether b I, II and 111 there were four male and three female dogs, 30 .6() Af d i under a stream f nitrogen, h three male and three female dogs, and three male and sodium hydride was a light gray solid. To 250 f two female dogs respectively. They were maintained on pure dimethylsulfoxide which had been flushed with the Same diet as he Control Group except that d1-(3,5- nitrogen, 4.8 g (0.2 mole) of the dry sodium hydride 3O di-tert-butyl-4-hydroxypheny fl was added I0 was add d, Thi Slurry was he t d l wl t 65 d the extent of 0.3, 1.0 and 3.0 weight per cent, based on held at this temperature for one hour. Hydrogen was the Weight of the g food, respectively The h liberuted as the dimethylsulfoxide anion was being Pound was added y dissolving it in the Wesson Oii P f d and a alm t l a l i lted, Aft to the limit of solubility and thereafter thoroughly mixcooling to 30, 21.2 g (0.05 mole) of (4,4- ing any additional amount of compound with the dog methylenebis-2,6-di-tert-butylpheno1) was added with food. The dogs had an average weight of 8 kilograms stirring. This reaction was exothermic and the temperaand ate an average of 200 grams of dog food per day. ture of the mixture was held below 60 during the addi- After 30 days, samples of plasma and serum were taken tion by using an ice bath for cooling and by using a slow and the serum analyzed for total lipid, total cholesterol, rate of addition. When the solid phenol contacted the free cholesterol and cholesterol-ester values. The resurface it turned a deep blue, however, on reacting sults are summarized in Table I.

TABLE 1 Serum Lipid Levels 30 Days No. Per Total Total Free Ester Group Dogs Cent"-" Lipids Cholesterol Cholesterol Cholesterol Control 6 694 120 3 89 Experimental l 7 0.3 465 56 12 44 Experimental ll 6 1.0 495 36 8 32 Experimental Ill 5 3.0 377 43 8 35 longer a gray solid separated. This mixture was heated with stirring for 1 hour at 60. The cooled mixture was then treated with 31.2 g (0.2 mole) of ethyl iodide. This addition took six minutes and an ice bath was needed to keep the temperature below After the mixture began to cool, it was heated at for one hour. The deep red color which formed on addition of the ethyl iodide disappeared after about 40 minutes of heating. The yellow mixture which remained formed a yellow solid on cooling. The mixture was treated with water and extracted with chloroform several times. The chloroform extracts were washed with water, dried with- 60 lyzed for cholesterol, phospholipid and triglyceride values. The results are summarized in Table [1.

TABLE ll Serum Lipid Values Days 0. Group Dogs Cholesterol Phospholipid Triglyceride Control 6 287 41 TABLE ll-Continued Serum Lipid Values'" 90 Days 0. Group Dogs Cholesterol Phospholipid Triglyceride Experimental 111 5 33 97 8 "Means values, expressed as Mg. 01' lipid/100 mlv of serum.

Residual serum from the dogs of the Control Group and Experimental Group Ill were separately pooled and the amount of aand B-lipoprotein in the serum for each group was determined by ultracentrifugation. The results are summarized in Table 111.

TABLE III Serum Lipoprotein Values 90 Days Group Total a-Lipoprotein Total BLipoprotein Control 355 1 39 Experimental 111 l 78 41 The compounds are administered internally and may be parenterally or orally administered, the latter being preferable. The compounds of this invention are generally solid at room temperature and for oral administration pharmaceutical preparations of this invention may be made following conventional techniques of the pharmaceutical chemist. These techniques involve granulating and compressing when necessary or variously mixing and dissolving or suspending the ingredients as appropriate to the desired end product. Numerous pharmaceutical forms to carry the compounds can be used. For example, the pure compound can be used or it can be mixed with a solid carrier. Generally, inorganic pharmaceutical carriers are preferable and particularly solid inorganic carriers.

In other tests which show effectiveness of the compositions and methods of this invention, test materials were administered daily to pure bred male beagle dogs. In some instances the test compound was given by capsule 125 mg/kg of body weight. In other instances the compound was admixed with the diet. The concentration of the test chemical in the diet was based on the dogs previous food consumption and provided approximately 125 mg/kg of body weight.

The test diets were prepared by dissolving or suspending the test compound in Mazola Corn Oil followed by heating to 9397C. for up to 30 minutes and mixing with the feed Wayne Dog Food, Krums, meal). All diets contained corn oil at the concentration of 5 percent by weight.

The dogs were fed ad libitum for one hour each day from pre-weighed food hoppers. Water was available ad libitum.

Results are summarized in the following table.

CHOLESTEROL VALUES IN MG/IOO ML SERUM Compound Compound Administration Administered In Capsules 1n oil mixed with Diet Weeks Weeks Com- Dog Baseline Values Baseline Avg. 2 4 6 1 2 3 4 pound A 1 144 153 182 171 162 128 151 155 A 2 141 127 121 122 128 116 151 120 B 3 121* 77 86 83 79 69 81 B 4 149* 153* 200 196 198 171 208 180 142 110 C 5 163 155 173 164 134 119 105 C 6 157 153 161 150 143 154 D 7 187 155 I76 155 140 157 D 8 221 220 212 134 140 157 E 9 153 141 142 145 137 122 121 105 63 E 10 197 192 186 192 192 183 165 I36 126 F 1 1 242 255 266 255 254 208 237 208 F 12 264 301 255 278 274 232 289 272 data not used in calculating averages Compound A 2,2'-methylenehis(6-tert-bulyl-4-chlorophenol) Compound 8 2,2-methylenehis( 6-tert-buty1-p-creso1) Compound C 4.4"methylenebisl2.6-bis(a-methylbenzyhphenol] Compound D 2-tert-hutyl-4.4methy1enedipheno1 Compound E 4,4-isopropy1idenebis( 2,6 di-tert-butylphenol) Compound F 4,4' methylenebis(2.6-di-sec-hutylpheno1) normal sterol was found, showing that the effect of the compound on the biosynthesis of cholesterol is most likely at an earlier stage than after the cyclization of squalene. Such effects are highly desirable, as the formation of abnormal sterols may produce untoward side effects.

Efficacious results are also obtained with other compounds of this invention and with other mammals such as cattle, sheep, rabbits, and others, and this invention is particularly desirable for treating humans.

For Compound F, the test diet was prepared by dissolving 80 grams ofthe compound in 920 grams of corn oil and mixing with 19 kg. of Wayne Dog Food, Krums.

No apparent reason was noted for the inconsistency of hypocholesterolemic response of dogs 5 and 6 for Compound C. However, for Compound D, results indicated that the dog with a greater hypocholesterolemia received an average dose of 106/mg/kg/day while dog 7 received an average dose of 81 mg/kg/day. Also, the dogs on Compound A rejected the test diet.

Dogs maintained on diets containing up to 3.0 weight per cent of di-( 3,5-di-tert-butyl-4- hydroxyphenyl)methane for 90 days have shown no adverse effect on growth or behavior. When sacrificed at ulant therapy. Two cases of pneumonia; one resulting in death in a severely ill elderly patient, occurred while patients were on drugs but are not considered to be related to drug administration.

administration of the drug with meals. No alteration in the hypochlolesterolemic potency was observed by giving the drug with meals. The appearance of periorbital edema was reported in two of the 13 patients. In one patient there was a probable potentiation of anticoagthe end of 90 days there were no gross or microscopic In the terminal case, the patient was a 69 year old pathology attributable to the di-( 3,5-di-tertbutyl-4- male with hypertension, generalized arteriosclerosis hydroxyphenyl)methane. The acute LD in rats is obliterans and right hemiplegia. Terminally, his SGOT greater than gms/kg. Rats fed on a diet containing was 135, the alkaline phosphatase Bodansky units, substantial amounts of di-(3,5-di-tert-butyl-4- prothrombin time 17/14 and an inversed A/G ratio. Kehydroxyphenyl)methane for 90 days suffered no 10 tosteroid excretion was increased from 7.7 mg/gm cregrossly harmful effects. There were no changes in proatinine on 1/26/65 to 18.6 mg/gm creatinine (termithrombin time, hematocrit, hemoglobin or white blood nal). Pneumonia is not an uncommon complication in counts. At 1.25 weight per cent of the diet there were elderly patients and is often the cause of death in hemino gross or histopathological changes. plegic individuals.

For the compound 4,4-ethylidenebis(2,6-di-tert- 15 The side effects and clinical course of patients in the butyl-phenol), the test diet, which provided a daily group of geriatric patients are noted above. A careful dose of approximately 125 i mg/kg, was given to toxicologic evaluation ofrenal, liver, bone marrow, and g (0 f r a 7 k p ri x p r 3 y after h adrenal function (ketosteroid and 17-oxycorticoid exend of the fourth Week- The test diet Was fed to g cretion) in these patients has failed to demonstrate abfor 4 weeks, control diet during the fifth week, and test 20 normalities definitely related to the drug. diet on Monday, Wednesday, and Friday during the Scrum lipid levels can be reduced by internally adsixth and seventh weeks with control diet on alternate ministering to a mammal a composition having a serum days. Results were as follows: lipid lowering amount of the galvanoxyl radical Cholesterol Values in mg/IOO ml Serum Average Baseline Dog No. l 2 3 4* 5 6 7 163 i 117 175 133 158 127 140 122 161 ii 128 164 94 76 111 152 104 '4 weeks plus 2 days After seven weeks, alkaline phosphatase levelson both t-C H tr-C 11 dogs were run and found to be elevated. Elevation of serum alkaline phosphatase was also noted on other s CH 0 dogs who had exhibited a hypocholesterolemic effect upon being treated with methylenebis-(2,6-di-tertt- 4 s afle butylphenol). Based on such evidence, itappears that i dogs treated i this P cholesterpl lowerwhere t-C H is a tert-butyl group. The galvanoxyl radimg may be associated w1th increase 1n alkallne phos- 40 cal may be referred to as 2,6-d1-tert-butyl-a-(3,5-d1- phatase, however, this observation has not been made in man. Thus, except for possibly one instance, there tert'butyl'4'0X0'25cyclohexadleml'yhdene)'p' have not been any cases of increases in alkaline phosl yfi t t th b 1 b l phatase during the term of administration of the drug d Z g est ar g e 2} E ma 6 g in man, even in the presence of significant lowering of Q an per.cem y Welg.t 0 t e compoun cholesterol and other lipids m xed with the diet, the followmg results were ob- There have been clinical studies using methylenebis(- filmed N 2,6-di-tert-butylphenol) in man. These studies show that reductions in serum lipids from the drug can be obtained in man using 0.1, 0.25, 0.5 and 1.0 grams of the ChOlesterol Values in mg/100 ml Serum compound per day. For example, in one group of 13 geriatric patients, the average initial total serum choles- Baseline Weeks on iet terol was 244 mg per cent in a control period extending Average 1 2 4 over six weeks. The drug caused a fall to a mean level A 54 122 [27 '27 of 181 mg per cent when the drug was given over a four 55 B 232 208 243 197 week period.

In another study with healthy volunteers receiving 0.1, 0.5 and 1.0 grams of drug per day over the four week period of drug administration, the average reduction in Serum Cholesterol eXPTeSSed in milligram P Compositions containing the galvanoxyl radicals are 7 100 m1 of Serum was and 35 respectivelyillustrated by those made by substituting the radical for All the side effects noted in the clinical studies were the active ingredient in Examples 1-1 1 which follow. in the group of 13 geriatric patients. Three patients The compounds are administered internally and may complained of epigastric burning, anorexia, nausea and be parenterally or orally administered, the latter being occasional vomiting. The symptoms were obviated by preferable. The compounds of this invention are generally solid at room temperature and for oral administration pharmaceutical preparations of this invention may be made following conventional techniques of the pharmaceutical chemist. These techniques involve granulating and compressing when necessary or variously mixing and dissolving or suspending the ingredients as appropriate to the desired end product. Numerous pharmaceutical forms to carry the compounds can be used. For example, the pure compound can be used or it can be mixed with a solid carrier. Generally, inorganic pharmaceutical carriers are preferable and particularly solid inorganic carriers. One reason for this is the large number of pharmaceutically acceptable inorganic materials which are known to be pharmaceutically safe and acceptable, as well as very convenient in preparing formulations. The compositions may take the form of tablets, linguets, powders, capsules, slurries, troches, or lozenges, prepared by standard pharmaceutical techniques. Tablet compositions may be coated or uncoated and they may be effervescent or noneffervescent. Conventional excipients for tablet formations may be used. For example, inert diluents, such as magnesium carbonate or lactose, disintegrating agents such as maize starch or alginic acid, and lubricating agents such as magnesium stearate may be used. A preferable tablet composition is one which comprises from about 10 to about 500 milligrams of di-(hydroxyphenyl)methane, particularly di-(3,5-di-tert-butyl-4- hydroxyphenyl)methane.

If a liquid carrier is used, the preparation may be in the form of a soft gelatin capsule, a syrup, a liquid solution or suspension. Most of the compounds of this invention are only slightly soluble in water, if at all, but are generally soluble in alcohols particularly glycols, to some extent. In general, those di-(hydroxyphenyl)methanes having greater hydrocarbon substitution are less soluble in water. Di-(3,5-di-tert-butyl-4- hydroxyphenyhmethane, for example, is insoluble in water, but is soluble in ethyl alcohol to the extent of about six weight per cent. The hydrocarbon solubility of most of the compounds of this invention is high enough to allow the use of various pharmaceutically acceptable oils as carriers. For example, vegetable or animal oils such as sunflower oil, safflower oil, maize oil or codliver oil can be used. Glycerine can also be used. With these latter solvents, from 25-30 per cent water may be added. When water alone is the carrier, the preparation can be administered in the form ofa slurry.

Emulsion compositions may be formulated using emulsifying agents such as sorbitan tri-oleate, polyoxyethylene sorbitan monooleate, lecithin, gum acacia or gum tragacanth. Aqueous based suspensions may be prepared with the aid of wetting agents such as polyethylene oxide condensation products of alkylphenols, fatty alcohols or fatty acids and with suspending agents, for example, a hydrophilic colloid such as polyvinylpyrrolidone. The emulsions and suspensions may contain conventional excipients such as sweetening agents, flowing agents, coloring materials and preservatives.

The compounds of this invention may be administered in the form ofa nutritive preparation in which the mixture of active ingredients is mixed with proteins, such as casein and carbohydrates. In addition to the active ingredients, dietary supplements such as vitamins, salts of glycerophosphoric acid, choline, inositol and amino acids such as methionine may be added.

The percentage of di-(hydroxyphenyl)methane in the pharmaceutical carrier may be varied. It is necessary that the di-(hydroxyphenyl)methane constitute a proportion such that a suitable dosage will be obtained. It is therefore preferred to use pharmaceutical compositions containing at least 10 weight per cent of the di-(hydroxyphenyl)methane. Activity increases with concentration of the agent in the carrier, but those compositions containing a significant amount of carrier, e.g. at least I per cent and preferably at least 5 per cent, are preferred as they allow for the easier administration of the di-(hydroxyphenyl)methane.

For parenteral use, the compounds of this invention can be formulated with sterile ingredients, compounds and packaged asceptically. They may be administered intravenously or intramuscularly. Useful solvents for formulation in such use are the polyhydric aliphatic alcohols and mixtures thereof. Especially satisfactory are the pharmaceutically acceptableglycols, such as propylene glycol, and mixtures thereof. Glycerine is another example of a polyol which is particularly useful. Up to 25-30 per cent by volume of water may be incorporated in the vehicle if desired. An per cent aqueous propylene glycol solution is a particularly convenient solvent system. A pH range, about 7.4, and isotonicity compatible with body isotonicity is desirable. Basicity may be controlled by addition of a base as required, and a particularly convenient base is monoethanolamine. It may often be desirable to incorporate a local anaesthetic and such are well known to those skilled in the art. For example, lidocaine (B-diethylamine-2,6- acetoxylidide, available from the Astra Chemical Company) may be employed at a level of up to about 20 mg/c.c., or even more.

Administration of the compounds of this invention by the oral route is preferred. Advantageous daily dosages can be as low as mg. for a human. Higher dosages can be used. Generally, one would choose to use the least amount which affords the desired magnitude of sought-after result. Such practice tends to minimize chances of appearance of undesirable side effects. A preferred range of daily dosage is about 0.25 to 7.5 grams. In terms of body weight, advantageous dosages are from 2.5 to 1,000 mg. per kg. of body weight per day with a preferred range of about 25 to I25 mg. per kg. of body weight per day. The daily dosage is preferably administered from one to four or five times daily in amounts of from about 150 mg. to about 2,000 mg, and these amounts are administered in dosage units containing at least 10 mg. of the di-(hydroxyphenyl)methane. For example, when administering the compound in tablet form several tablets containing from, say 10 to 25 mg. of active compound can be administered, up to 4 or more times daily. Alternatively, larger dosage units containing more of the di( hydroxyphenyl)methane, say 50 to 500 mg. can be administered at less frequent intervals.

For parenteral applications daily dosages of about one-tenth of that used for oral treatment are advantageous. Thus, daily dosages can be as low as 15 mg. for a human, i.e. 0.25 mg. per kg. of body weight. A convenient upper limit is about 6 grams. From about 5 to about 1000 mg. per injection (dosage unit) in concentrations of about 5 to 200 mg/c.c., with from I to 3 injections of from 1 c.c. to 10 c.c. daily will give the required amount. Preferred formulations will contain from 50 to about 150 mg./c.c. to be given in one injection of from 1 c.c. to 5 c.c.

The oral or parenteral dose may be individually determined by the physcician or veterinarian. Larger or smaller doses can be used and, in some cases, might be preferred in individual cases. Likewise, administration need not be on a daily basis, although this is preferred,

EXAMPLE 1 Di( 3,S-di-tert-butyI-4-hydroxyphenyl)methane is compressed into tablets of 100 mg. each which can be administered orally as antihypercholesterolemic agents.

EXAMPLEZ 300 Mg. of 4,4-ethylidenebis(2,6-di-tertbutylphenol) are filled into a No. 2 hard gelatin capsule.

EXAMPLE 3 50 Gms. of 4,4"isopropylidenebis(2,6-di-tertbutylphenol), gms. of calcium sulfate and 25 gms. of sucrose are thoroughly mixed and granulated with hot 10 per cent gelatin solution. The wetted mass is passed through a No. l6 U.S. standard mesh screen directly onto drying trays. The granules are dried at I20F. and passed through a No. 20 U.S. standard mesh screen. These granules are then mixed with gms. starch, 5 gms. talc and 3 gms. stearic acid, passed through a No. 60 U.S. standard mesh screen and then compressed into tablets containing 150 mg. of active ingredient.

EXAMPLE 4 75 Mg. of 2-tert-butyl-4,4'-methylenediphenol are mixed with 225 mg. of peanut oil to a thick slurry and filled into a soft gelatin capsule.

EXAMPLE 5 Linguets are obtained by combining the following components:

4.4'-mcthylcncdi-2.6-xylcnol, mg. 25 Lactose, mg. 50 Confcctioncrs sugar. mg. 60 Stearin, mg. 2 Talc, mg. I3

EXAMPLE 6 To a mixture of 140 gms. of 2,2'-methylenebis(6- tert-butyI-4-chlorophenol) and 33.7 gms. of corn oil are added 3 gms. of gum acacia and 1.5 gms. of gum tragacanth. To the thoroughly triturated mixture is added slowly with stirring a solution of 0.1 gms. of a cetyl alcohol polyoxyethylene condensate, 40 gms. of can sugar, 0.03 gms. of propyl-p-hydroxybenzoate, 0.3 gms. of methyl-p-hydroxybenzoate, 0.002 g. of edible dyestuff and 1 10 gms. of water. After the incorporation of a suitable flavoring agent, such as imitation wild cherry, the mixture is homogenized by passage through a conventional homogenizer and there is thus obtained an emulsion suitable for oral administration in accordance with the present invention. The emulsion is bottled in half-pint bottles.

I0 Gms. of

methylbenzyl)-phenol] are dissolved in a mixture of 83 grrns. of water, 250 gms. glycerol and I25 gms. of ethyl alcohol. To the resultant solution is added to a solution of 300 gms. of sucrose and 150 gms. of water. By the incorporation of a suitable flavoring agent and coloring material there is obtained a syrup suitable for oral administration in accordance with the present invention.

EXAMPLE 8 25 Gms. of sodium glycerophosphate, 25 gms. of calcium glycerophosphate and mgs. of 4,4- methylenebis(2,6-di-sec-butylphenol) are intimately mixed. The mixture is added gradually to 900 gms. of soluble casein in a conventional mixer and mixed until homogenous. There is thus obtained a dietary supplement suitable for oral administration in accordance with this invention.

EXAMPLE 9 An intimate mixture is prepared with conventional mixing equipment of 3 gms. of pyridoxine hydrochloride, 100 gms. of nicotinic acid, 100 gms. of nicotinamide, 5 gms. of methionine, 15 gms. of choline bitartrate, I50 mgs. of ascorbic acid, 5 gms. of calcium pantothenate, 10 gms. of riboflavin and 1,000 gms of bis(- 3,5-ditert-butyl-4-ethoxyphenyl)methane. The mixture is filled into capsules which are then suitable for oral administration in accordance with this invention.

EXAMPLE I0 382 Gms. of propylene glycol are agitated for one hour while saturating with nitrogen gas. 12.4 Gms. of 4,4'-methylenebis (2,6-di-tert-butylphenol) are then added and the mixture is stirred for 30 minutes more. Then 95 c.c. of nitrogen saturated water is slowly added. After 5 minutes of further stirring 7.85 c.c. of monoethanolamine is added. Throughout the addition of ingredients the temperature is maintained below 30C. by cooling as required. Solution is completed by stirring under nitrogen. The resulting 500 c.c. of clear, light-colored solution contains approximately 50 mg./c.c. of 4,4-methylenebis-(2,6-di-tert-butylphenol) and is slightly alkaline. It is subdivided and samples are filled under nitrogen into 10 c.c. multi-dose vials sealed with butyl rubber stoppers. 3 CC. doses can be withdrawn in the standard manner, by piercing the stopper with a hypodermic needle, injecting air and withdrawing the solution into the syringe. In this manner paren- Water to make 400 c.c.s

The above solution contains mgs./c.c. of 4,4- methylenebis-(2,6-di-tert-butylphenol) and can be administered intramuscularly in I c.c. doses, once a day, giving a daily dosage of 150 mgs.

In general, solid dosage forms contain from 5 to 60 per cent of active ingredient while liquid dosage forms usually contain 1 to 20 per cent active ingredient. Greater or lesser amounts can be employed.

Thus, preferred embodiments of this invention are: A method of treating a mammal whose serum lipid level is to be reduced which comprises internally administering to said mammal a serum lipid-lowering amount of a compound having the formula a b R d R R l HO C H c R R wherein R" and R" are tert-butyl,

R and R are alkyl of l to 4 carbons, and R and R are independently selected from hydrogen or methyl.

This method may be used in treating a mammal with elevated or abnormally high lipid levels, as well as those having normal amounts of lipid.

Such method can comprise internally administering to a mammal the following compounds of the above formula (II).

di-( 3 ,5-di-tert-butyl-4-hydroxyphenyl )methane 4,4 -isopropylidenebis( 2,6-di-tert-butylphenol) 4,4'-ethylidenebis( 2,6-di-tert-butylphenol) 4,4 '-methylenebis( 2 ,6-d i-sec-butylphenol) Also, this invention comprises the preferred methods of treating a mammal whose serum lipid level is to be reduced which comprises internally administering to said mammal a serum lipid-lowering amount of [2,6-bis-( a-methylbenzyl )phenol 2-tert-butyl-4,4'-methylenediphenol,

2,2 '-methylenebis( 6-tert-butyl-4-chlorophenol), 2,2'-methylenebis( fi-tert-butyl-p-cresol), or

bis( 3 ,5di-tert-butyl-4-ethoxyphenyl )methane.

In another preferred embodiment, I provide therapeutic compositions effective in lowering serum lipid levels in mammals which comprise from 5 to 500 milligrams of a compound of Formula IV and a significant amount of a pharmaceutical carrier. Thus, for example, I provide such compositions where said compound is di-( 3,5-di-tert-butyl-4-hydroxyphenyl)methane. Also, I provide such compositions having at least weight per cent of a compound of Formula IV and at least 5 per cent by weight of a pharmaceutically acceptable carrier. These compositions may be in capsule form or tablet form. For example, such capsules may contain the compound di-(3,5-di-tert-butyl-4- hydroxyphenyl)methane.

Also, there is provided therapeutic compositions having at least l0 weight per cent of a compound of Formula IV and at least 5 per cent by weight of a pharmaceutically acceptable carrier,

:1. wherein R" in said formula is methyl. b. wherein R" in said formula is methyl,

c. wherein R" in said formula is hydrogen,

d. wherein R" in said formula is hydrogen,

e. wherein R and R in said formula are methyl,

f. wherein R and R in said formula are butyl,

g. wherein the compound is 4,4'-isopropylidenebis(2,6-

di-tert-butylphenol) h. wherein the compound is 4,4'-ethylidenebis(2,6-ditert-butylphenol), and

i. wherein the compound is 4,4-methylenebis(2,6-disec-butylphenol).

Also, there is provided therapeutic compositions having at least 5 per cent by weight of a pharmaceutically acceptable carrier and at least 10 weight per cent of a compound such as di-(3,5-di-tert-butyl-4-hydroxyphenyl)methane 4,4'-methylenebis[2,6-bis(a-methylbenzyUphenol] 2,2-methylenebis(6-tert-butyl-4-chlorophenol) bis( 3 ,5-di-tert-butyl-4-ethoxyphenyl )methane 2-tertbutyl-4,4'-methylenediphenol, or 2,2'-methylene(6-tert-butyl-p-cresol).

The therapeutic compositions discussed above are preferably in unit dosage form. Typical dosage forms are exemplified by Examples l-ll above.

Above, when clinical testing was discussed, a case was mentioned of a possible increase in alkaline phosphatase in man. This case was a healthy volunteer who showed an increase after receiving for 4 weeks l.0 gram daily of methylenebis(2,6-di-tert-butylphenol). However, repeat determinations of alkaline phosphatase were normal, and the subject remained asymptomatic.

I claim:

I. A method of treating a mammal whose serum lipid level is to be reduced which comprises internally administering to said mammal, in unit dosage form a serum lipid-lowering amount of a compound having the formula wherein R and R" are tert-butyl,

R and R are alkyl of l to 4 carbons, and

R and R are independently selected from hydrogen or methyl.

2. The method of claim 1 wherein said compound is di-(3,5-di-tert-butyl-4-hydroxyphenyl)methane.

3. A method of treating a dog whose plasma lipid level is to be reduced, which comprises internally administering from about 0.25 mg. to about 1.0 gram of di-( 3,5-di-tert-butyl-4-hydroxyphenyl)methane per kilogram of body weight per day.

4. The method of claim I wherein R is methyl.

5. The method of claim 4 wherein R is methyl.

6. The method of claim 1 wherein R is hydrogen.

7. The method of claim 6 wherein R" is hydrogen. 8. The method of claim I wherein R and R are methyl.

9. The method of claim 1 wherein R and R are butyl.

10. The method of claim 1 wherein said compound is 4,4-isopropylidenebis( 2,6-di-tert-butylphenol).

11. A method of treating a mammal, in unit dosage form whose serum lipid level is to be reduced which comprises internally administering to said mammal a serum lipid-lowering amount of 4,4'-methylene-bis- [2,6-bis-(a-methylbenzyl)phenol].

12. A method of treating a mammal, in unit dosage form whose serum lipid level is to be reduced which comprises internally administering to said mammal a serum lipid-lowering amount of 2,2-methylene-bis(6- tert-butyl-4-chlorophenol).

13. A method of treating a mammal, in unit dosage form whose serum lipid level is to be reduced which comprises internally administering to said mammal a lipid-lowering amount of bis( 3,5-di-tert-butyl-4- ethoxyphenyl)methane.

14. A method of treating a mammal, in unit dosage form whose serum lipid level is to be reduced which comprises internally administering to said mammal a serum lipid-lowering amount of 2-tert-butyl-4,4'- methylenediphenol.

15. A method of treating a mammal, in unit dosage form whose serum lipid level is to be reduced which comprises internally administering to said mammal a serum lipid-lowering amount of 2,2'-methylene-bis(6- tert-butyl-p-cresol).

16. A therapeutic composition in unit dosage form having at least 10 per cent by weight of 4,4- methylenebis[2,6-bis(a-methylbenzyl)phenol] with at least 5 per cent by weight of a pharmaceutically acceptable carrier;

17. A therapeutic composition in unit dosage form having at least 10 per cent by weight of Z-tert-butyl- 4,4'-methylenediphenol with at least 5 per cent by weight of a pharmaceutically acceptable carrier.

18. A method of treating a mammal whose serum lipid level is to be reduced which comprises internally administering to said mammal, in unit dosage form, a serum lipid-lowering amount of 4,4-ethylidenebis( 2,6 di-tert-butylphenol).

19. A method of treating a mammal whose serum lipid level is to be reduced which comprises internally administering to said mammal, in unit dosage form, a serum lipid-lowering amount of 4,4-methylenebis(2,6- di-sec-butylphenol).

20. The method of claim 1 wherein said compound is 4,4-methylene bis(2-ethyl-6-tert-butylphenol).

21. The method of claim 1 wherein said compound is 4,4-methylene bis(2-isopropyl-6-tert-butylphenol).

22. The method of claim 6 in which R is the same as R 23. The method of claim 7 in which R is the same as R. 

1. A METHOD OF TREATING A MAMMAL WHOSE SERUM LIPID LEVEL IS TO BE REDUCED WHICH COMPRISES INTERNALLY ADMINISTERING TO SAID MAMMAL, IN UNIT DOSAGE FORM A SERUM LIPID-LOWERING AMOUNT OF A COMPOUND HAVING THE FORMULA
 2. The method of claim 1 wherein said compound is di-(3,5-di-tert-butyl-4-hydroxyphenyl)methane.
 3. A method of treating a dog whose plasma lipid level is to be reduced, which comprises internally administering from about 0.25 mg. to about 1.0 gram of di-(3,5-di-tert-butyl-4-hydroxyphenyl)methane per kilogram of body weight per day.
 4. The method of claim 1 wherein Rd is methyl.
 5. The method of claim 4 wherein Re is methyl.
 6. The method of claim 1 wherein Rd is hydrogen.
 7. The method of claim 6 wherein Re is hydrogen.
 8. The method of claim 1 wherein R and Rc are methyl.
 9. The method of claim 1 wherein R and Rc are butyl.
 10. The method of claim 1 wherein said compound is 4,4''-isopropylidenebis(2,6-di-tert-butylphenol).
 11. A method of treating a mammal, in unit dosage form whose serum lipid level is to be reduced which comprises internally administering to said mammal a serum lipid-lowering amount of 4, 4''-methylene-bis-(2,6-bis-( Alpha -methylbenzyl)phenol).
 12. A method of treating a mammal, in unit dosage form whose serum lipid level is to be reduced which comprises internally administering to said mammal a serum lipid-lowering amount of 2, 2''-methylene-bis(6-tert-butyl-4-chlorophenol).
 13. A method of treating a mammal, in unit dosage form whose serum lipid level is to be reduced which comprises internally administering to said mammal a lipid-lowering amount of bis(3,5-di-tert-butyl-4-etHoxyphenyl)methane.
 14. A method of treating a mammal, in unit dosage form whose serum lipid level is to be reduced which comprises internally administering to said mammal a serum lipid-lowering amount of 2-tert-butyl-4,4''-methylenediphenol.
 15. A method of treating a mammal, in unit dosage form whose serum lipid level is to be reduced which comprises internally administering to said mammal a serum lipid-lowering amount of 2, 2''-methylene-bis(6-tert-butyl-p-cresol).
 16. A therapeutic composition in unit dosage form having at least 10 per cent by weight of 4,4''-methylenebis(2,6-bis( Alpha -methylbenzyl)phenol) with at least 5 per cent by weight of a pharmaceutically acceptable carrier.
 17. A therapeutic composition in unit dosage form having at least 10 per cent by weight of 2-tert-butyl-4,4''-methylenediphenol with at least 5 per cent by weight of a pharmaceutically acceptable carrier.
 18. A method of treating a mammal whose serum lipid level is to be reduced which comprises internally administering to said mammal, in unit dosage form, a serum lipid-lowering amount of 4, 4''-ethylidenebis(2,6-di-tert-butylphenol).
 19. A method of treating a mammal whose serum lipid level is to be reduced which comprises internally administering to said mammal, in unit dosage form, a serum lipid-lowering amount of 4, 4''-methylenebis(2,6-di-sec-butylphenol).
 20. The method of claim 1 wherein said compound is 4,4''-methylene bis(2-ethyl-6-tert-butylphenol).
 21. The method of claim 1 wherein said compound is 4,4''-methylene bis(2-isopropyl-6-tert-butylphenol).
 22. The method of claim 6 in which R is the same as Rc.
 23. The method of claim 7 in which R is the same as Rc. 